11{62 ,15{62 -PGE{HD 2{B ,15{62 -PGF{HD 2{301 {B , and 15{62 -PGF{HD 2{331 {B , 15-acetates methyl esters

ABSTRACT

Novel methods are disclosed for transforming PGA2 and 15 Beta PGA2 and their acetates, methyl esters, and acetate methyl esters to various prostanoic acids and esters of the PGE2 and PGF2 series. Some of the latter are novel and are useful for the same pharmacological purposes as PGE2 and PGF2 .

United States Patent Pike et al.

113,15 B-PGE,,I5 B-PGF AND 15 B-PGF IS-ACETATES METHYL ESTERS Inventors: John E. Pike; William P. Schneider,

both of Kalamazoo, Mich.

Assignee: The Upjohn Company, Kalamazoo,

Mich.

Filed: Oct. 3, 1973 Appl. No.: 403,091

Related U.S. Applimtion Data Division of Ser. No. |59,478, July 2, 197l, Pat. No.

3,772,350, which is a continuation-in-part of Ser. No. H.390, Sept. I l, I970. abandoned.

US. CL. 260/488 R; 260/348 A; 260/3485 L; 260/448.8 R; 260/468 D; 260/491; 260/499; 260/514 D; 424/305; 424/311 Int. Cl. C07C 177/00 Dec. 23, 1975 [58] Field of Search 260/438 R. 514 D, 468 D [56] References Cited UNITED STATES PATENTS 3,706,789 l2/l972 Bergstrom et 260/488 R 3,795,697 3/1974 Bergstrom et al 260/488 R Primary Examiner-Vivian Garner Attorney, Agent, or FirmMorris L. Nielsen [57] ABSTRACT 4 Claims, No Drawings IIBJSB-PGEQJS ror ,Aivnls -pcn IS-ACETA ES METHYL ESTERS CROSS REFERENCE TO RELATED APPLICATIONS This is a division of application Ser. No. 159,478, filed July 2, 1971, which is a continuatiomin'part of our co-pending application Ser. No. 71,390, filed Sept. I], 1970.

DESCRIPTION OF THE INVENTION WM H l aam/awrweo Some of the derivatives of prostanoic acid are known as prostaglandins. One of those, prostaglandin E (PGE has the following formula:

'WCOO Another, prostaglandin F (PGF has the formula:

Still another, prostaglandin F (PGF has the formula:

In Formulas l to IV and in the formulas recited hereinafter in the specification and claims, broken line attachments to the cyclopentane ring indicate substituents in alpha configuration, i.e., below the plane of the cyclopentane ring. Heavy solid line attachments to the cyclopentane ring indicate substituents in beta configuration, i.e., above the plane of the cyclopentane ring.

The side-chain hydroxy at C-lS in Formulas II to IV is in S (alpha) configuration. That configuration is shown by attachment of said side-chain hydroxy to C- l 5 with a dotted line and hydrogen with a heavy solid line. The alternative configuration for the side-chain hydroxy at 015 is known as R or epi (beta), and is 2 shown when necessary at attachment of said side-chain hydroxy to C15 with a heavy solid line and hydrogen with a dotted line, thus The prostaglandin corresponding to PGE (Formula II) but with the R or epi configuration at C-IS will be designated ISB-PGE- See Nature, 212, 38 I966) for discussion of the stereochemistry of the prostaglandins.

These conventions regarding formulas, names, and symbols for derivatives of prostanoic acid apply to the formulas, names and symbols given hereinafter in the specification and claims. When reference is made hereinafter to the compounds of Formulas II to IV, by the symbols PGE PGF- or PGF or to the methyl esters of any of those, l5(S) configuration will be intended and by established custom, S or alpha will not be mentioned in the name or symbol. For all of the other compounds recited hereinafter, the configuration at C-l5 will be identified in the name as 15B whenever the l5(R) configuration is intended.

Molecules of the known prostaglandins each have several centers of asymmetry, and can exist in racemic (optically inactive) form and in either of the two enan tiomeric (optically active) forms, i.e., the dextrorotatory and levorotatory forms. As drawn, Formulas II to IV each represent the particular optically active form of the prostaglandin which is obtained from certain mammalian tissues, for example, sheep vesicular glands, swine lung, or human seminal plasma, or by carbonyl and/or double bond reduction of a prostaglandin so obtained. See, for example, Bergstrom et al., Pharmacol. Rev. 20, l ([968) and references cited therein.

The several aspects of this invention relate to novel methods for preparing PGE PGF and PGF- their acetates and methyl esters, and the I5B-epimers of those compounds, to novel intermediates used in those methods, to novel methods used to make those intermediates, and to certain novel and pharmacologically useful analogs of PGE PGF and PGF The novel and pharmacologically useful F GE PGFQQ and the formulas:

analogs of this invention have In Formulas V, VI, and VII, R is hydrogen, alkyl of one to 8 carbon atoms, inclusive, cycloalkyl of 3 to carbon atoms, inclusive, aralkyl of 7 to 12 carbon atoms, inclusive, phenyl, or phenyl substituted with one to 3 chloro or alkyl of one to 4 carbon atoms, inclusive. Also encompassed by Formulas V, VI, and VII are pharmacologically acceptable salts when R, is hydrogen. In Formulas V and VII, Y is H OH H OH H t! or ln Formula V, B is H" \)H or H, on.

In Formula Vl, indicates attachment to the ring in alpha or beta configuration.

It will be observed that each of the novel compounds of Formulas V and VI has a hydroxy group attached to the l l-position in beta configuration. In PGE PGF and PGF and in the compounds of For mula VII, the hydroxy at C-ll is attached in alpha configuration.

With regard to Formulas V, VI, and VII, examples of alkyl of one to 8 carbon atoms, inclusive, are methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, and isomeric forms thereof. Examples of cycloalkyl of 3 to 10 carbon atoms, inclusive, which includes alkyl-sub stituted cycloalkyl, are cyclopropyl, 2-methylcyclopro' pyl, 2.3-dimethyl cyclopropyl, 2,3-diethylcyclopropyl, Z-butylcyclopropyl, cyclobutyl, 2-methylcyclobutyl, 3-propylcyclobutyl, 2,3,4-triethylcyclobutyl, cyclopentyl, 2,2-dimethylcyclopentyl, 3-pentylcyclopentyl, 3- tert-butylcyclopentyl, cyclohexyl, 4-tert-butylcyclohexyl, 3-isopropylcyclohexyl, 2,2-dimethylcyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and cyclodecyl. Examples of aralkyl of 7 to l2 carbon atoms, inclusive, are benzyl, phenethyl, lphenylethyl, 2 phenylpropyl, 4-phenylbutyl, 3-phenylbutyl, 2-( lnaphthylethyl), and l-(2-naphthylmethyl). Examples of phenyl substituted by one to 3 chloro or alkyl of one to 4 carbon atoms, inclusive, are p-chlorophenyl, mchlorophenyl, o-chlorophenyl, 2,4-dichlorophenyl, 2,4,6-trichlorophenyl, p-tolyl, m-tolyl, o-tolyl, p-ethylphenyl, ptert-butylphenyl, 2,5-dimethylphenyl, 4- chloro-Z-methylphenyl, and 2,4-dichloro-3methylphen l.

The known prostanoic acid derivatives, PGE PGF- and PGF- and their esters and pharmacologically acceptable salts are extremely potent in causing various biological responses. For that reason, these compounds are useful for pharmacological purposes. See, for example, Bergstrom et al., cited above, and references cited therein. A few of those biological responses are systemic arterial blood pressure lowering in the case of the POE- and PGE compounds as measured, for example, in anesthetized (pentobarbital sodium) pentolinium-treated rats with indwelling aortic and right heart cannulas; pressor activity, similarly measured, for the PGF compound; stimulation of smooth muscle as shown, for example, by tests on strips of guinea pig ileum, rabbit duodenum, or gerbil colon; potentiation of other smooth muscle stimulants; antilipolytic activity as shown by antagonism of epinephrine-induced mobilization of free fatty acids or inhibition of the spontaneous release of glycerol from iso lated rat fat pads; inhibition of gastric secretion in the case of the PGE compounds as shown in dogs with secretion stimulated by food or histamine infusion; activity on the central nervous system; controlling spasm and facilitating breathing in asthmatic conditions; decrease of blood platelet adhesiveness as shown by platelet-to-glass adhesiveness, and inhibition of blood platelet aggregation and thrombus formation induced by various physical stimuli, e.g., arterial injury, and various biochemical stimuli, e.g., ADP, ATP, serotonin, thrombin, and collagen; and in the case of the PGE compounds, stimulation of epidermal proliferation and keratinization as shown when applied in culture to embryonic chick and rat skin segments.

Because of these biological responses, these known prostaglandins are useful to study, prevent, control or alleviate a wide variety of diseases and undesirable physiological conditions in birds and mammals, including humans useful domestic animals, pets, and zoological specimens, and in laboratory animals, for example, mice, rats, rabbits, and monkeys.

For example, these compounds, and especially the PGE compounds, are useful in mammals, including man, as nasal decongestants. For this purpose, the compounds are used in a dose range of about 10 pg. to about l0 mg. per ml. of a pharmacologically suitable liquid vehicle or as an aerosol spray, both for topical application.

The PGE and PGF compounds are useful in the treatment of asthma. For example, these compounds are useful bronchodilators or as inhibitors of mediators, such as SRS-A and Histamine which are released from cells activated by an antigemantibody complex. Thus, these compounds control spasm and facilitate breathing in conditions such as bronchial asthma, bronchitis, bronchiectasis, pneumonia and emphysema. For these purposes, these compounds are administered in a variety of dosage forms, e.g., orally in the form of tablets, capsules, or liquids; rectally in the form of tablets, capsules, or liquids; rectally in the form of suppositories; parenterally, subcutaneously, or intramuscularly, with intravenous administration being preferred in emergency situations; by inhalation in the form of aerosols or solutions for nebulizers; or by insufflation in the form of powder. Doses in the range of about ().()l to 5 mg. per kg. of body weight are used I to 4 times a day, the exact dose depending on the age, weight, and condition of the patient and on the frequency and route of administration. For the above use these prostaglandins can be combined advantageously with other antiasthmatic agents, such as sympathomimetics (isoproterenol, phenylephrine, ephedrine, etc); xanthine derivatives (theophylline and aminophyllin); and corticosteroids (ACTH and predinisolone). Regarding use of these compounds see South African Patent No. 681,055.

The POE: compounds are useful in mammals, including man and certain useful animals, e.g., dogs and pigs, to reduce and control excessive gastric secretion, thereby reducing or avoiding gastrointestinal ulcer formation, and accelerating the healing of such ulgers already present in the gastrointestinal tract. For this purpose, the compounds are injected or infused intravenously, subcutaneously, or intramuscularly in an infusion dose range about 0.1 pg. to about 500 pg. per kg. of body weight per minute, or in a total daily dose by injection or infusion in the range about 0.1 to about 5 20 mg. per kg. of body weight per day, the exact dose depending on the age, weight, and condition of the patient or animal, and on the frequency and route of administration.

The PGE PGF and PGB compounds are useful whenever it is desired to inhibit platelet aggregation, to reduce the adhesive character of platelets, and to remove or prevent the formation of thrombi in mammals, including man, rabbits, and rats. For example, these compounds are useful in the treatment and prevention of myocardial infarcts, to treat and prevent postoperative thrombosis, to promote patency of vacular grafts following surgery, and to treat conditions such as atherosclerosis, arteriosclerosis, blood clotting defects due to lipemia, and other clinical conditions in which the underlying etiology is associates with lipid imbalance or hyperlipidemia. For these purposes, these compounds are administered systemically, e. g., intravenously; subcutaneously, intramuscularly, and in the fonn of sterile implants for prolonged action. For rapid response, especially in emergency situations, the intravenous route of administration is preferred. Doses in the range about 0.005 to about 20 mg. per kg. of body weight per day are used, the exact dose depending on the age, weight, and condition of the patient or animal, and on the frequency and route of administration.

The PGE,, PGF and PGF compounds are especially useful as additives to blood, blood products, blood substitutes, and other fluids which are used in artificial extracorporeal circulation and perfusion of isolated body portions, e.g., limbs and organs, whether attached to the original body, detached and being preserved or prepared for transplant, or attached to a new body, During these circulations and perfusions, aggregated platelets tend to block the blood vessels and portions of the circulation apparatus. This blocking is avoided by the presence of these compounds. For this purpose, the compound is added gradually or in single or multiple portions to the circulating blood, to the blood of the donor animal, to the perfused body portion, attached or detached to the recipient, or to two or all of those at a total steady state does of about 0.00l to I mg. per liter of circulating fluid. It is especially useful to use these compounds in laboratory animals, e.g., cats, dogs, rabbits, monkeys, and rats, for these purposes in order to develop new methods and techniques for organ and limb transplants.

The PGE compounds are extremely potent in causing stimulation of smooth muscle, and are also highly active in potentiating other known smooth muscle stimulators, for example, oxytocic agents, e.g., oxytocin, and the various ergot alkaloids including derivatives and analogs thereof. Therefore PGE for example, is useful in place of or in combination with less than usual amounts of these known smooth muscle stimulators, for example, to relieve the symptoms of paralytic ileus, or to control or prevent atonic uterine bleeding after abortion or delivery, to aid in expulsion of the placenta, and during the puerperium. For the latter purpose, the PO5 compound is administered by intravenous infusion immediately after abortion or delivery at a dose in the range about 0.0! to about 50 ,ug. per kg. of body weight per minute until the desired effect is obtained. Subsequent doses are given by intravenous, subcutaneous, or intramuscular injection or infusion during puerperium in the range 0.01 to 2 mg. per kg. of body weight per day, the exact dose depending on the age, weight. and condition of the patient or animal.

The PGE and PGF compounds are useful as hypotensive agents to reduce blood pressure in mammals, including man. For this purpose, the compounds are administered by intravenous infusion at the rate about 0.0l to about 50 pg. per kg. of body weight per minute, or in single or multiple doses of about 25 to 500 ug. per kg. of body weight total per day.

The PS5,, PGA and PGF compounds also in crease the flow of blood in the mammalian kidney, thereby increasing volumeand electrolyte content of the urine. Therefore, these compounds are useful in managing cases of renal disfunction, especially those involving blockage of the renal vascular bed. lllustra tively, the compounds are useful to alleviate and correct cases of edema resulting, for example, from massive surface burns, and in the management of shock. For these purposes, the compounds are preferably first administered by intravenous injection at a dose in the range l0 to I000 pg. per kg. of body weight or by intravenous infusion at a dose in the range 0.1 to 20 pg. per kg. of body weight per minute until the desired effect is obtained. Subsequent doses are given by intravenous, intramuscular, or subcutaneous injection or infusion in the range 0.05 to 2 mg. per kg. of body weight per day. t

The P05 PGF and PGF compounds are useful in place of oxytocin to induce labor in pregnant female animals, including man, cows, sheep, and pigs, at or near term, or in pregnant animals with intrauterine death of the fetus from about 20 weeks to term. For this purpose, the compound is infused intravenously at a dose 0.0l to 50 pg. per kg. of body weight per minute until or near the termination of the second stage of labor, i.e., expulsion of the fetus. These compounds are especially useful ,when the female is one or more weeks post-mature and natural labor has not started, or 12 to hours after the membranes have ruptured and natural labor has not yet started.

The PGF- PGF, and PGE, compounds are useful for controlling file reproductive cycle in ovulating female mammals, including humans and animals such as monkeys, rats, rabbits, dogs, cattle, and the like. For that purpose, P05 or PGF for example, is administered systemically, e.g., intravenously, subcutaneously, and intravaginally, at a dose level in the range 0.00l mg. to about 20 mg. per kg. of body weight of the female mammal, advantageously during a span of time starting approximately at the time of ovulation and ending approximately at the next expected time of menses or just prior to that time. Additionally, expulsion of an embryo or fetus (abortion) is accomplished by similar administration of the compound during the first third of the normal mammalian gestation period.

As mentioned above, the POE compounds are po tent antagonists of epinephrine-induced mobilization of free fatty acids. For this reason, this compound is useful in experimental medicine for both in vitro and in vivo studies in mammals, including man, rabbits and rats, intended to lead ,to the understanding, prevention, symptom alleviation, and cure of diseases involving abnormal lipid mobilization and high free fatty acid levels, e.g., diabetes mellitus, vascular diseases, and hyperthyroidism.

The novel Formula-V, -Vl, and -Vll PGE PGF and PGF analogs of this invention each cause the biological responses described above for PGE PGF and PGF respectively, and each of these novel compounds is accordingly useful for the above described corresponding purposes, and is used for those purposes in the same manner as described above.

PGE PGF and PGF and their esters and pharmacologically acceptable salts are all potent in causing multiple biological responses even at low doses. For example, PGE is extremely potent in caus ing vasodepression and smooth muscle stimulation, and is also potent as an antilipolytic agent. Moreover, for many applications, these known prostaglandins have an inconveniently short duration of biological activity. In striking contrast, the novel analogs of Formulas V, VI, and Vll are substantially more specific with regard to potency in causing prostaglandin-like biological responses, and have a substantially longer duration of biological activity. Therefore, each of these novel prostaglandin analogs is surprisingly and unexpectedly more useful than one of the corresponding above-mentioned known prostaglandins for at least one of the pharmacological purposes because it has a different and narrower spectrum of biological activity than the known prostaglandins, and therefore is more specific in its activity and causes smaller and fewer undesired side effects than when the known prostaglandin is used for the same purpose. Moreover, because of its prolonged activity, fewer and smalle doses of the novel prostaglandin analog can frequently be used to attain the desired result.

The novel Formula-V, -Vl, and -Vll prostaglandin analogs are used as described above in free acid form in ester form. or in pharmacologically acceptable salt form. When the ester form is used, the alkyl esters are preferred, especially the alkyl esters wherein the alkyl moiety contains one to 4 carbon atoms, inclusive. Of those alkyl, methyl and ethyl are especially preferred for optimum absorption of the compound by the body or experimental animal system.

Pharrnacologically acceptable salts of these prostaglandin analogs useful for the purposes described above are those with pharmacologically acceptable metal cations, ammonium, amine cations, or quarternary ammonium cations.

Especially preferred metal cations are those derived from the alkali metals, e.g., lithium, sodium and potassium, and from the alkaline earth metals, e.g., magnesium and calcium, although cationic forms of other metals, e.g., aluminum, zinc and iron, are within the scope of this invention.

Pharrnacologically acceptable amine cations are those derived from primary, secondary, or tertiary amines. Examples of suitable amines are methylamine, dimethylamine, trimethylamine, ethylamine, dibutylamine, triisopropylamine, N-methylhexylamine, decylamine, dodecylamine, allylamine, crotylamine, cyclopentylamine, dicyclohexylamine, benzylamine, dibenzylamine, a-phenylethylamine, B-phenylethylamine, ethylenediamine, diethylenetriamine, and like aliphatic, cycloaliphatic, and araliphatic amines containing up to and including about 18 carbon atoms, as well as heterocyclic amines, e.g., piperidine, morpholine, pyrrolidine, piperazine, and lower-alkyl derivatives thereof, e.g., l-methylpiperidine, 4-ethylmorpholine, l-isopropylpyrrolidine, Z-methylpyrrolidine, l,4- dimethylpiperazine, Z-methylpiperidine, and the like, as well as amines containing water-solubilizing or by drophilic groups, e.g., mono-, di-, and triethanolamine, ethyldiethanolamine, N-butylethanolamine, 2-aminol-butanol, 2-amino-2-ethyll ,3-propanediol, 2amino- Z-methyll -propanol, tris( hydroxymethyl )aminometharie, N-phenylethanolamine, N-(p-tert-amylphenyl)- diethanolamine, galactamine, N-methylglucamine, N- methylglucosamine, ephedrine, phenylephrine, epi nephrine, procaine, and the like.

Examples of suitable pharmacologically acceptable quaternary ammonium cations are tetramethylammonium, tetraethylammonium, benzyltrimethylammonium, phenyltriethylammonium, and the like.

As discussed above, these novel prostaglandin analogs are administered in various ways for various purposes; e.g., intravenously, intramuscularly, subcutaneously, orally, intravaginally, rectally, buccally, sublingually, topically, and in the form of sterile implants for prolonged action.

For intravenous injection or infusion, sterile aqueous isotonic solutions are preferred. For that purpose, it is preferred because of increased water solubility to use the free acid form or the pharmacologically acceptable salt form. For subcutaneous or intramuscular injection, sterile solutions or suspensions of the acid, salt, or ester form in aqueous or non-aqueous media are used. Tablets, capsules, and liquid preparations such as syrups, elixirs, and simple solutions, with the usual pharmaceutical carriers are used for oral or sublingual administra tion. For rectal or vaginal administration, suppositories, tampons, ring devices, and preparations adapted to generate sprays or foams or to be used for lavage, all prepared as known in the art, are used. For tissue implants, a sterile table or silicone rubber capsule or other object containing or impregnated with the substance is used.

The novel compounds of Formulas V, VI, and Vll wherein R is other than hydrogen, i.e., the esters wherein R, is alkyl of one to 8 carbon atoms, inclusive, cycloalkyl of 3 to 10 carbon atoms, inclusive, aralkyl of 7 to 12 carbon Eatoms, inclusive, phenyl, or phenyl substituted with one to 3 chloro or alkyl of one to 4 carbon atoms, inclusive, are prepared from the corresponding acids of Formulas V, VI, and Vll, i.e., wherein R, is hydrogen, by methods known in the art. For example, the alkyl, cycloalkyl, and aralkyl esters are prepared by interaction of said acids with the appropriate diazohydrocarbon. For example, when diazomethane is used, the methyl esters are produced. Similar use of diazoethane, diazobutane, l-diazo-2-ethylhexane, diazocyclohexane, and phenyldiazomethane, for example, gives the ethyl, butyl, Z-ethylhexyl, cyclohexyl, and benzyl esters, respectively.

Esterification with diazohydrocarbons is carried out by mixing a solution of the diazohydrocarbon in a suitable inert solvent, preferably diethyl ether, with the acid reactant, advantageously in the same or a different inert diluent. After the esterification reaction is complete, the solvent is removed by evaporation, and the ester purified if desired by conventional methods, preferably by chromatography. It is preferred that contact of the acid reactants with the diazohydrocarbon be no longer than necessary to effect the desired esterification, preferably about one to about ten minutes, to avoid undesired molecular changes. Diazohydrocarbons are known in the art or can be prepared by methods known in the art. See, for example, Organic Reactions, John Wiley & Sons, lnc., New York, N.Y., Vol. 8, pp. 389-394 I954).

An alternative method for esterification of the carboxyl moiety of the novel PGF-type or POE-type compounds of Formulas V, VI, and Vll comprises transformation of the free acid to the corresponding silver salt,

9 followed by interaction of that salt with an alkyl iodide. Examples of suitable iodides arc methyl iodide, ethyl iodide, butyl iodide, isobutyl iodide, tert-butyl iodide, cyclopropyl iodide, cyclopentyl iodide, benzyl iodide, phenethyl iodide, and the like. The silver salts are prepared by conventional methods, for example, by dissolving the acid in cold dilute aqueous ammonia, evaporating the excess ammonia at reduced pressure, and then adding the stoichiometric amount of silver nitrate.

The phenyl and substituted phenyl esters of the Formula-V, -Vl, and Vll compounds are prepared by silylating the acid to protect the hydroxy groups, for example, replacing each OH with OSi(CH Doing that may also change COOH to COOSi- (CH;,) A brief treatment of the silylated compound with water will change COO-Si(CH back to COOH, Procedures for this silylation are known in the art and are discussed hereinafter. Then, treatment of the silylated compound with oxalyl chloride gives the acid chloride which is reacted with phenol or the appropriate substituted phenol to give a silylated phenyl or substituted phenyl ester. Then the silyl groups, e.g., OSi(Cl-l;,) are changed back to OH by treatment with dilute acetic acid. Procedures for these transformations are known in the art.

The novel Formula-V, Vl, and VII acids (R is hydrogen) are transformed to pharmacologically acceptable salts by neutralization with appropriate amounts of the corresponding inorganic or organic base, examples of which correspond to the cations and amines listed above. These transformations are carried out by a variety of procedures known in the art to be generally useful for the preparation of inorganic, i.e., metal or ammonium salts, amine acid addition salts, and quaternary ammonium salts. The choice of procedure depends in part upon the solubility characteristics of the particular salt to be prepared. in the case of the inorganic salts, it is usually suitable to dissolve the acid in water containing the stoichiometric amount of a hydroxide, carbonate, or bicarbonate corresponding to the inorganic salt desired. For example, such use of sodium hydroxide, sodium carbonate, or sodium bicar bonate gives a solution of the sodium salt of the pros tanoic acid derivative. Evaporation of the water or addition of a water-miscible solvent of moderate polarity, for example, a lower alkanol or a lower alkanone, gives the solid inorganic salt if that form is desired.

To produce an amine salt, the acid is dissolved in a suitable solvent of ether moderate or low polarity. Ex amples of the former are ethanol, acetone, and ethyl acetate. Examples of the latter are diethyl ether and benzene. At least a stoichiometric amount of the amine corresponding to the desired cation is then added to that solution. If the resulting salt does not precipitate, it is usually obtained in solid form by addition of a miscible diluent of low polarity or by evaporation. If the amine is relatively volatile, any excess can easily be removed by evaporation. It is preferred to use stoichio metric amounts of the less volatile amines.

Salts wherein the cation is quaternary ammonium are produced by mixing the acid with the stoichiometric amount of the corresponding quaternary ammonium hydroxide in water solution, followed by evaporation of the water.

The novel compounds of Formulas V, VI, and VII wherein R is hydrogen or methyl, i.e., the free acids and the methyl esters, and also PGE PGF and PGF and the methyl esters of those are prepared 10 by novel methods which are described hereinafter. For those methods, one of the following starting materials is used:

W M COGRZ VI Ia N H" 0R H0 WCQQRZ Vlll V WOORZ IX ln Formulas Vlla, VIII, and IX, R is either hydrogen or methyl, and R is hydrogen or acetyl.

It will be observed that the compounds encompassed by Formula Vlla are also encompassed by Vll. Thus, some Formula-VII compounds are useful both as intermediates and for pharmacological purposes.

These Formula-Vlla, Vlll, and IX starting materials are all derivatives of prostanoic acid. The compounds of Formula Vlll are known in the art or are available by methods known in the art. See, for example, Weinheimer et al., Tetrahedron Letters, No. 59, 5 l (1969); H. W. Youngken, .lr. (ed), Food-Drugs from the Sea, Proc. Marine Technology Society, pp. 3l 1-314 (1969). The Formula-VlIl compound wherein R and R are both hydrogen is designated lSB-PGA- alternatively l5(R)-PGA or l5-epi-PGA.,. The other compounds encompassed by Formula Vlll are designated l5B-PGA acetate, l5filPGA methyl ester, and 15,8-PGA acetate methyl ester.

The compounds of Formula Vlla are new in the art and methods for preparing them are described hereinafter. The Formula-Vlla compound wherein R and R are both hydrogen is designated l5BPGE alternatively l5(R)-PGE or l5-epi-PGE The other compounds encompassed by Formula Vlla are designated l5B-PGE lfi-acetate, ISB-PGE methyl ester, and l5B-PGE acetate methyl ester.

The compounds of Formula IX are known in the art. See, for example, British Specification 1,097,533. Novel methods for preparing these Formula-IX compounds are described hereinafter. The FormulalX compound wherein R and R; are hydrogen is designated PGA The other compounds encompassed by Formula IX are designated PGA acetate, PGA methyl ester, and PGA acetate methyl ester.

All of the compounds of Formulas Vlla, VIII, and [X are obtained by extraction from a marine invertebrate. The compounds of Formula Vlla and VIII, i.e., the 158 compounds, are obtained from colonies of Plexauru Iwmumalla (Esper), 1792, forma R. The compounds of Formula IX, i.e., the 15(8) or alpha compounds, are obtained from colonies of Plexaura homomalla (Esper), I792, forma S.

These Plexaura lzomomalla forms are members of the subclass Octocorallia, order Gorgonacea, suborder Holaxonia, family Plexauridae, genus Plexaura. See, for example, Bayer, The Shallow-Water Octocorallia of the West Indian Region", Martinus Nijhoff, The Hague (I96l Colonies of these Plexuuru homomalla forms are abundant on the ocean reefs in the zone from the low-tide line to about 25 fathoms in the tropical and subtropical regions of the western part of the Atlantic Ocean, from Bermuda to the reefs of Brazil, including the eastern shore reefs of Florida, the Caribbean island and mainland reefs, and the Gulf of Mexico island and mainland reefs. These colonies are bush-like or small tree-like in habit, and are readily identified for collection as Plexaura homomalla (Esper), 1792, by those of ordinary skill in this art. Forms R and S are distin guished by the methods described in Preparation 1 below.

The colonies of these two forms of Plexaura homomalla are easily separated into an outer bark-like cor tex and an inner wiry proteinaceous stem or skeleton. Symbiotic algae or Zooxanthellae are also present in the colonies. Weinheimer et al., cited above, disclose the occurence of the Formula-VIII compounds wherein R and R are both hydrogen and wherein R is methyl and R is acetyl in the air dried cortex of Plexaura homomalla (Esper)".

The choice of isolation or extraction method is deter mined by the particular Formula-VIIa, -VIII, or -IX compound desired. Maximum yield of the Formula- VIII or -IX diester is realized by freezing whole or coarsely cut or chopped fresh Plexaura homomalla colonies within an hour and preferably sooner after the colonies are removed from the reef. For small scale collections, this freezing is done advantageously by contacting the colonies or pieces with solid carbon dioxide. For larger scale collections, other suitable freezing methods are known to the art. The frozen colonies or colony pieces should be kept frozen, preferably below about 20 C. until the extraction takes place.

The major component of fresh Plexaura homomalla (Esper), I792, forma R is lSfi-PGA acetate methyl ester, the Formula-VIII compound wherein R is methyl and R is acetyl. Relatively minor components are the hydroxy methyl ester, the acetate, and the hydroxy acid of Formula VIII and the ISB-PGE compounds encompassed by Formula VIIa. Of the latter, the l5B-PGE acetate methyl ester (R is methyl and R is acetyl) is the most abundant. The major component of Plexaura homomalla (Esper), I792, forma S is PGA acetate methyl ester, the Formula-IX compound wherein R is methyl and R is acetyl. Relatively minor components are the hydroxy methyl ester, the acetate, and the hydroxy acid of Formula IX, and the PGE compounds corresponding to Formula VIIa but having the (5) configuration.

When the acetate methyl ester compound of Formula VIIa, VIII, or IX (R is methyl, R is acetyl) is desired as a starting material, a suitable method comprises grinding the frozen whole Plexuuru homomalla colonies or colony pieces, advantageously in a hogger to a particle size with the largest dimension about 5 mm., and then extracting the resulting particles with any of the usual organic solvents, preferably one with moderate to high polarity, e.g., dichloromethane or methanol, advantageously, for 15 to 30 minutes in a high speed mixer. The desired compounds are isolated from the extract by evaporation, and then chromatography of the resulting residue. By this procedure, about 24 g. of I5B-PGA acetate methyl ester, and about one g. each of ISB-PGA: methyl ester and ISB-PGE are obtained by dichloromethane extraction of I500 g. of frozen 12 Plexaura homomalla (Esper), I792, forma R colonies or colony pieces. Similarly, relatively large amounts of PGA acetate methyl ester are obtained from frozen Plexaura homomalla (Esper), I792, forma S.

When the IS-hydroxy methyl ester of Formula VIIa, VIII, or IX (R is methyl, R is hydrogen) is desired as a starting material, a suitable method comprises grinding the frozen whole Plexaura homomalla colonies or colony pieces as above, and then contacting the resulting particles with a lower alkanol, preferably methanol or ethanol, at 25 C. for several days. The solvent is then evaporated and the residue chromatographed to give substantially larger amounts of the hydroxy methyl ester compound relative to the acetate methyl ester compound. When the contact between the Plexaura homomalla particles and the alkanol is substantially shorter, substantially the same amount and ratio of the various Formula-VIIa, -VIII, or -IX compounds is obtained with the alkanol as with dichloromethane. An alternative method for obtaining these IS-hydroxy methyl esters is described below.

When l5B-PGA l5B-PGE or PGA (R and R in Formulas VIIa, VIII, and IX are both hydrogen) are desired as starting materials in the novel processes of this invention, they are prepared from the corresponding methyl esters and l5-acetate methyl esters after those have been extracted from the Plexaura homomalla colonies or colony pieces as described above. A suitable method for removing the acetyl group of each of the Formula-VIIa, VIII, and -IX IS-acetate methyl esters comprises mixing the acetate methyl ester in lower alkanol solution, preferably in methanol solution, with a strong acid, e.g.. perchloric acid, for about 15 hours at 25 C. A suitable method for removing the methyl group of any ofthe Formula-VIIa, -VIII, and -IX methyl esters is the enzymatic hydrolysis described in West Germany Offenlegungschrift No. 1,937,9I2, reprinted in Farmdoc Complete Specifications, Book No. 14, No. 6869R, Week R March 18, I970.

Another method for obtaining l5B-PGA ISB-PGE or PGA from Plexuura homomalla colonies or colony pieces comprises freezing the Plexaura homomalla colonies or colony pieces, preferably at a temperature below about 20 C., and then allowing the colonies or colony pieces to thaw and warm to a temperature in the range 20 to 30 C. The thawed colonies or colony pieces are then maintained in the range 20 to 30 C. for at least 24 hours. After that treatment, substantially none of the Formula-VIIa, -VIII, and -IX compounds wherein R is methyl and R is acetyl are present, the principal Formula-VIIa, -VIII, and -IX compounds present being those wherein R and R are both hydrogen, the minor components being those wherein R is methyl and R is hydrogen or wherein R is hydrogen and R is acetyl. As before, Formula-Vlla and Vlll compounds are obtained from colonies of Plemura homomalla (Esper), I792, forma R, and Formula-IX compounds are obtained from colonies of Plcxaura homomalla (Esper), I792. forma S.

A preferred procedure for the PGA and PGE type free acids comprises grinding the Plexaura homomalla colonies or colony pieces, preferably to a particle size with the largest dimension about 5 mm., and then main taining the mixture in contact with water at a temperature in the range 20 to 30 C. for at least 24 hours. This mixture is filtered. and the filtrate is extracted with an appropriate water-immiscible solvent, eg, ethyl acetate. The solid residue is also extracted with an appro- 13 priate solvent, e.g, methanol. The two extracts are evaporated, and the total residue is chromatographed to give Formula-Vila and -VIII or Formula-IX compounds, the principal component in each case being the compound wherein R and R; are both hydrogen.

Since our invention of the novel processes for trans forming PGA and lSB-PGA and their methyl esters and acetate methyl esters to the various prostanoic acids and esters disclosed herein, it has now been found that small amounts of the 5,6-trans compounds of PGA and IB-PGA and their methyl esters and acetate methyl esters are also obtained from Plexaura homomalla (Esper), 1792, forms R and S. These 5,6- Trans compounds are extracted with and accompany the corresponding PGA -type compounds through many of their transformations. For example, PGA containing 5,6-trans-PGA yields a mixture of PGE and 5,6-trans-PGE by the process represented in Chart E below.

When it is desired, for pharmacological purposes, to prepare the major products of this invention free of 5,6-trans compounds, those 5,6-trans compounds are separated either from the starting reactants or from the products. In either case, several methods are available for separating the 5,6-trans-PG compounds from the PG compounds. One method is by means of a silversaturated ion-exchange resin (for example, see E. A. Emken et al., J. Am. Oil Chemists Soc. 41, 388 1964)), illustrated below in Preparations 5 and 6. The other method is by preferentially forming a mercuric acetate adduct of the 5,6-cis compound which is extractable into polar solvents illustrated below in Preparation 7.

Following the processes discussed herein and the procedures of the Examples below, the 5,6-transPG (and -l5B-PG compounds are transformed to other 5,6-trans-PG (and -BPG compounds, e.g., 5,6- trans-PGA to 5,6-trans-PGE 5,6-ti'ans-l5,8-F'GA acetate methyl ester to 5,6-trans-l5fi-PGF acetate methyl ester, and the like.

As mentioned above, the FormuIa-VIIa, -VIII, and -IX compounds are starting materials for the preparation of PGE PGF and PGF the methyl esters of those, and also the novel compounds of Formulas V and VI, and some of the novel compounds of Formula VII. The novel processes using these starting materials will now be described.

The Formula -VIII and -IX starting materials are both of the PGAtype. According to the novel processes of this invention, those are first transformed to cprresponding PGE-type compounds. The chemical reactions involved in those transformations are shown generically in Chart A.

In Chart A, R is hydrogen, methyl, or Si(A) wherein A is alkyl of one to 4 carbon atoms, inclusive, aralkyl of 7 to 12 carbon atoms, inclusive, phenyl, or phenyl substituted with one or 2 fluoro, chloro, or alkyl of one to 4 carbon atoms, inclusive; G is wherein R is hydrogen, acetyl, or Si(A);, when R, is hydrogen or methyl, and R is -Si(A);; when R, is Si(A) R is hydrogen or methyl; and B is 14 H OH or H OH wherein R is as defined above, and Z is wherein A is as defined above In Formula XI, of Chart A, indicates attachment of the epoxy oxygen to the ring in alpha or beta configuration. In Formulas XII and XIII of Chart A, indicates attachment of hydroxy to the ring in alpha or beta configuration.

CHART A COOR4 x (0x idati on} \ZMOQRZ (reduction) 'WCQORZ XI I (hydrolysis) COORZ XI I H0 It will be observed in Chart A that the Formula-Xll and Xlll products each encompass four stereoisomeric groups of compounds. Included are compounds with the l 1a,15(S) configuration of PGE (Formula 11, above compounds with the configuration of l 101,153- PGE: (Formula Vlla) as Obtained from Plexaum Immunzullu (Espcr), 1792, forma R, and both the 15(8) and l5(R) compounds with the 113 configuration of the novel Formula V compounds of this invention wherein Y is i.e., 1 lpl-PGE and llB,15B-PGE4 If the Formula-X11 or (lll product is to have the 15(S) configuration, e.g., PGE. or llfi-PGE then the Formula-X starting material should have the 15(8) configuration, i.e., G

should be 11' a 1513 compound of Formula X1] or Xlll is desired, e.g., l5B-PGE or 1 1,8, ISB-PGE then the Formula-X starting material should have the l5(R) or l5-epi configuration. i.e., G should be H OR As described above, Formula-1X starting materials wherein R is hydrogen or methyl and R is hydrogen, and with the 15(S) configuration. are obtained from Plemura liomvmulla (Esper), 1792, forma S. Those same compounds are also produced by reacting the corresponding l5(R) (beta) compound with a hydrocarbyl or halohydrocarbyl sulfonyl chloride or bromide, preferably a lower alkylsulfonyl chloride or bromide, especially methanesulfonyl chloride or bromide, or a benzeneor substituted-benzenesulfonyl chloride or bromide, e.g., p-toluenesulfonyl chloride. This reaction is done in the presence of at least sufficient tertiary amine, e.g., triethylamine, to absorb the hydrogen chloride or hydrogen bromide by-product, and at a low temperature, preferably in the range 1 5 to +15 C. The presence of an inert liquid diluent, e.g., tetrahydrofuran, is helpful to maintain a mobile homogeneous reaction mixturev At 0 C. and with methanesulfonyl chloride, usually to 60 minutes is a sufficient reaction time. The product is hydrolyzed to a mixture of 15(5) (alpha) and l5(R) hydroxy compounds. These are separated by procedures known in the art, and the 15 (s) product is purified by procedures known in the art. advantageously by chromatography on silica gel. This reaction is also used to transform 15(5) Formula- 1X starting materials wherein R is hydrogen or methyl and R is hydrogen to the corresponding l5(R) compounds. In each case, a mixture of l5(R) product and 15(8) starting material is obtained, the components of which are separated as described above.

Another method of transforming a ISB-PG compound to a PG compound is by converting it to a mix* 16 ture of PG l5formate and l5/3PG liformillc conr pounds, separating the PG lS-formate, and hydrolyzing the PG 15-formate to the desired PG compound (see J.E. Pike et al., J. Org. Chem. 34. 3552 (1969)).

The mixture of alpha and beta lS-formates is prepared by maintaining the 158 compound, cg. 15,8- PGE lSB-PGA or ISBPGF in formic acid buffered with an alkali metal formate in the range 10 to 50 C. until a substantial amount of the 15/3 compound, eg. l5/3-PGE l5formate, has been transformed to the PG l5formate, The mixture of the PG l5-formate and 15B IS-formates thus obtained is then separated by known methods, e.g. by chromatography.

The PG l5 formate can then he hydrolyzed to the desired PG IS-hydroxy compound. The ISB-PG l5- formate yields the corresponding lSB-PG IS-hydroxy compound, which is then recycled through the above steps for further isomerization to the PG compound if desired.

This procedure is also useful to transform a PG compound to a 153 PG compound, by obtaining a mixture of the intermediate PG lS-formate and ISB-PG l5-formate compounds, separating them, and hydrolyzing them to the respective PG lfi-hydroxy and [SB-PG l5-hydroxy compounds. In this case, the PG compound is recycled for further isomerization to the 15B com pound.

Referring again to Chart A, the transformation of starting material X to epoxide X1 is carried out by reacting X with any agent known to epoxidizc an alt unsaturated ketone without reacting with isolated carbon-carbon double bonds, for example see Steroid Reactions, Carl Djerassi, ed., Holden-Day Inc., 1963, p. 593. Especially preferred are aqueous hydrogen peroxide or an organic tertiary hydroperoxide. See, for example, Organic Peroxides, A.V. Tobolsky et al, In terscience Publishers, N.Y., 1954. For this purpose, the peroxide or hydroperoxide is employed in an amount of at least one equivalent per mole of Formula-X reactant in the presence of a strong base, e.g., an alkali metal hydroxide, a metal alkoxide, or a quaternary ammonium hydroxide. For example, there is employed 1ithium hydroxide, sodium hydroxide, potassium hydrox ide, lithium ethoxide, lithium octyloxide, magnesium methoxide, magnesium isopropoxide, benzyltrimethylammonium hydroxide, tetraethylammonium hy droxide, butyltrimethylammonium hydroxide, butyldiethylphenylammonium hydroxide, benzylethyldimethylammonium hydroxide, benzyldimcthyloctadecylammonium hydroxide, benzyldodecyldime thylammonium hydroxide, decyldimethylphenylain monium hydroxide, and the like. See, for example, Sidgwick, Organic Chemistry of Nitrogen, Third Edi-- tion, rev. by Miller and Spring-all, Oxford, 1966, pp. 1 16-127.

The ratio of alpha to beta epoxide formed in the reaction is related to four factors: the epoxidizing agent, the base, the diluent, and the temperature. Hy drogen peroxide is employed in the concentrations usually available, for example 3% to although 30% is especially convenient. When the alpha epoxide is the desired product, tert-butyl hydroperoxide is espe' cially preferred as the epoxidizing agent. Examples of other organic tertiary hydroperoxides useful for this purpose are tert-pentyl hydroperoxide, decahy dronaphthyl hydroperoxide, mar-dimethylbenzyl hydroperoxide, and 1,1-diphenylethyl hydroperoxide. The base is present in the proportion of 0.1-3.0, prefer ably about U. IU.5 equivalent of base per mole of starting material X when R is methyl and R is acetyl', preferably about 1.5-2.5 equivalent of base per mole of starting material VIII or IX wherein R and R are hydrogen. When the alpha epoxide is the desired product, lithium hydroxide, lithium or magnesium alkoxides of one to 8 carbon atoms, and benzyltrimethylammonium hydroxide are the preferred bases, although the lithium and magnesium compounds are especially preferred.

It is advantageous to use an inert liquid diluent in the epoxidation step to produce a mobile homogenous reaction mixture, for example, a lower alkanol, dioxane, tetrahydrofuran, dimethoxyethane, dimethylsulfoxide, or dimethylsulfone. When the alpha epoxide is preferred, tetrahydrofuran or the less polar dimethoxyethane are especially preferred as the diluent. A reaction temperature in the range 60O to 0 C. is generally preferred, especially below l() C. The lower temperatures below 30 C. are especially preferred for favoring formation of alpha epoxide over beta epoxide. At a temperature of C., the epoxidation is usually complete in 3 to 6 hours. It is also preferred that the reaction be carried out in an atmosphere of an inert gas, e.g., nitrogen, helium, or argon. When the reaction is complete as shown by the absence of starting material on TLC plates (3% acetone in dichloromethane), the reaction mixture is neutralized, and the epoxy product is isolated by procedures known in the art, for example, evaporation of the diluent and extraction of the residue with an appropriate water-immiscible solvent, e.g., ethyl acetate.

This transformation of X to XI usually produces a mixture of Formula-XI alpha and beta epoxides both with either the IS( R) or lS(S) configuration depending on the configuration at C-l5 in the Formula-X starting material. Although these mixtures are separated into the individual alpha and beta isomers, for example, by chromatography by procedures known to be useful for separating alpha and beta epoxide mixtures, it is usually advantageous to transform the Formula-XI mixture of alpha and beta epoxides to the corresponding mixture of Formula-XII [Ia and IIB hydroxy compounds. The latter mixture is then readily separated into the Ila and [I3 compounds, for esample, by chromatography of silica gel].

During the transformation of epoxides XI to alcohols XII, an alpha epoxide yields an Ila-hydroxy compound, and a beta epoxide yields an IIB-hydroxy compound. The ratio of alpha to beta epoxides in XI, and hence the eventual ratio of Ila and IIB alcohols in XII, produced from starting material X is dependent in large measure on the nature of R in X. Recall that G is defined as H OR or H OR wherein R is hydrogen, acetyl, or -Si(A):; wherein A is as defined above. For either definition of G, i.e., R configuration or S configuration, when R, is hydrogen, more Formula-Xi beta epoxide is formed than when R is acetyl, and more Formula-XI beta epoxide is formed when R, is acetyl than when R, is -Si-(A);,. For example, when G in formula X is H OH.

18 the preferred basic hydrogen peroxide epoxidation gives about equal amounts of alpha and beta epoxides, but when G in formula X is about 3 parts of alpha epoxide and one part of beta epoxide are obtained, and when G in formula X is about 4 parts of alpha epoxide and one part of beta epoxide are obtained, both reactions with the same epoxidation reagent. When G in formula X is H OH.

about one part of alpha epoxide and 3 parts of beta epoxide are obtained with basic hydrogen peroxide, but when G is about 6 parts of alpha epoxide and 4 parts of beta epoxide are obtained with the same epoxidation reagent.

Each of the novel Formula-V and -VI compounds of this invention has a hydroxy attached in beta configuration to the cyclopentane ring. Some of the compounds of Formula V, i.e., when Y is are encompassed by Formula XII (Chart A). The other Formula-V compounds and all of the Formula VI compounds are prepared as described below from Formula- XII compounds wherein the hydroxy at C4] is attached in beta configuration. Therefore, when a Formula-V or Formula-VI compound is described as a final product for pharmacological purposes, there is advantage in choosing a corresponding Formula-X starting material which gives the maximum amount of beta epoxide during the transformation of X to XI. Those will be the Formula-X compounds wherein G is H OH or H on.

On the other hand, when a prostanoic acid product with the natural lloz configuration for the hydroxy at C-ll is the desired final product, e.g., PGE PGF or PGF there is advantage in choosing a Formula- X starting material which gives a greater amount of the alpha epoxide during the transformation of X to XI. Those would be Formula-X compounds wherein X is or the corresponding 15(5) compounds.

As mentioned above, the starting materials of Formula X encompass not only the Formula-VIII and -IX compounds obtained from plexaura homomalla but also the silyl compounds of Formula X,,. When desired as reactants, these silyl compounds are prepared by silylation of PGA ISBPGA- or the methyl esters of those. These silylations are carried out by procedures known in the art. See, for example, Pierce, Silylation of Organic Compounds", Pierce Chemical Co., Rockford, Ill. (1968). The C-IS hydroxy group of PGA I5,B- PGA or their methyl esters is transformed to an O-Si(A) moiety wherein A is as defined above, sufficient silylating agent being used according to known procedures to accomplish that. The necessary silylating agents for this purpose are known in the art or are prepared by methods known in the art. See, for example, Post, Silicones and Other Organic Silicon Compounds, Reinhold Publishing Corp, New York, NY. (I949). In the case of PGA and lSB-PGA excess silylating agent and prolonged treatment also transform the COOH to COOSi(A);,. It is optional in transforming X to XI whether or not this COOl-l of PGA or l5B-PGA is esterified to COO-Si(A);,, since that ester group is transformed to COOH during formation and isolation of the Formula-XI epoxide product.

The various A's ofa Si(A) are alike or different. For example, an Si(A) can be trimethylsilyl, dimethylphenylsilyl, or methylphenylbenzylsilyl.

When it is desired to retain the Si-(A) moiety at C-l5 in the Formula-XI epoxide product, for example, to give steric control in a subsequent reaction, it is important in isolating the epoxide that the presence of acid be avoided and that contact with water be minimized unless the water is kept cold, i.e., below about l C.

Referring again to Chart A, the transformation of epoxide XI to hydroxy compound XII is accomplished by reduction with chromium (II) salts, e.g., chromium (II) chloride or chromium (II) acetate. Those salts are prepared by methods known in the art, e.g., Inorganic Syntheses, VIII, I25 (I966); ibid., VI, 144 (1960); ibid. III, I48 ([950); ibid. I, 122 (1939); and references cited in those. This reduction is carried out by procedures known in the art for using chromium (II) salts to reduce epoxides of aB-unsaturated ketones to ,B-hydroxy ketones. See, for example, Cole et al., J. Org. Chem. 19, I31 (1954), and Neher et al., Helv. Chem. Acta 42, I32 (I959). In these reactions, the absence of air and strong acids is desirable. If it is desired to maintain a -Si(A) moiety on C-l5, a neutral reaction mixture is preferred. An especially preferred procedure is to generate the chromium (II) ion in the presence of the Formula-XI epoxide, for example, by mixing the epoxide with a chromium (III) salt, e.g., the chloride, with metallic zinc in the presence of acetic acid. The desired Formula-XII compound is isolated from the reduction reaction mixture by methods known in the art, care being taken to minimize contact of the product with acid and water, especially warm water. when retention of a -Si(/\):; at C-l is desired.

Unexpectedly, amalgamated aluminum metal has also been found to be useful as a reducing agent in place of chromium (II) salts to transform Formula XI epoxides to Formula XII hydroxy compounds. This reagent was previously not known to be useful for this type of reaction. This use of amalgamated aluminum 20 represents a distinct and separate aspect of this invention.

Amalgamated aluminum is prepared by procedures known in the art, for example, by contacting aluminum metal in the form of foil, thin sheet, turnings, or granules with a mercury (II) salt, for example, mercuric chloride, advantageously in the presence of sufficient water to dissolve the mercury (II) salt. Preferably, the surface of the aluminum metal is free of oxide. That is readily accomplished by physical removal of the usual oxide layer, e.g., by abrasion or scraping, or chemically, e.g., by etching with aqueous sodium hydroxide solution. It is only necessary that the aluminum surface be amalgamated. The amalgamated aluminum should be freshly prepared, and maintained in the absence of air and moisture until used.

The reductive opening of the Formula-XI epoxide ring is accomplished by contacting said epoxide with the amalgamated aluminum in the presence of a hydroxylic solvent and sufficient inert organic liquid dilu ent to give a mobile and homogeneous reaction mixture with respect to the hydroxylic solvent and said epoxide. Among hydroxylic solvents, water is especially preferred although lower alkanols, e.g., methanol and ethanols are also operable.

Examples of inert organic liquid diluents are normally liquid ethers such as diethyl ether, tetrahydrofuran, dimethoxyethane, diglyme (dimethyl ether or diethylene glycol), and the like. Especially preferred is tetrahydrofuran. When a water-immiscible liquid dilu ent is used, a mixture of water and methanol or ethanol is especially useful in this reaction since the latter two solvents also aid in forming the desired homogeneous reaction mixture. For example, a mixture of diethyl ether and water is used with sufficient methanol to give a homogeneous reaction mixture.

This reductive opening requires two hydrogen atoms per molecule of epoxide. Amalgamated aluminum reacts readily with water and more slowly with other hydroxylic solvents to give hydrogen. One atomic equivalent of aluminum required 3 molecular equivalents of the hydroxylic solvent to give 3 atomic equivalents of hydrogen. Therefore, one molecular equivalent of epoxide requires two-thirds atomic equivalent of aluminum and two molecular equivalents of the hydroxylic solvent. Evolution of hydrogen gas (H molecules) is observed during this reductive opening of the epoxide. It is not known whether the reductive opening is caused by hydrogen atoms or hydrogen molecules. However, some of the hydrogen gas escapes from the reaction mixture. Therefore, it is preferred to use an excess of amalgamated aluminum and hydroxylic solvent, preferably at least one atomic equivalent of aluminum and three molecular equivalents of hydroxylic solvent per molecular equivalent of epoxide. Because of the relatively high economic value of the epoxide compared with amalgamated aluminum and hydroxylic solvents, it is preferred to assure maximum yields of the desired Formula-XII hydroxy compound by use of substantially greater excess of amalgamated aluminum and hydroxylic solvent, e.g., up to ten times or more of those reagents than is theoretically required.

The reductive opening of the epoxide is carried out by mixing a solution of the epoxide in the organic diluent with the amalgamated aluminum and the hydroxylic solvent. Since the reaction is exothermic, it is usually advantageous to cool the solution to a low temperature, e.g., 2() C. to 0 C., before adding the amal gamated aluminum and hydroxylic solvent and to maintain the reaction mixture in the range 20 to 30 C. by external cooling. This is especially advantageous when water is used as the hydroxylic solvent. Higher reaction temperatures are operable but not preferred when a high yield of the Formula-XII products is desired. Stirring is preferred during the reaction since the reaction mixture is heterogeneous with respect to the solution and the amalgamated aluminum.

For reasons not understood, better yields and a shorter reaction time are usually observed when only part of the amalgamated aluminum is added at the start of the reaction, additional portions being added during the reaction, e.g., at one-hour intervals, than when the entire amount of amalgamated aluminum is added at the start of the reaction. A suitable procedure is to add about one-third of the amalgamated aluminum at the start, about one-third after I hour, and another third after a second hour. The course of the reaction is ad vantageously determined by withdrawing small portions of the solution and determining the presence or absence of starting material by thin layer chromatography. For example, when R is methyl and G is H ococH Xla wherein G and M are as defined above and R is a cation of an alkali or alkaline earth metal or a quaternary ammonium group.

Thus, the Formula-XI epoxide compound is treated with a hydroxide or oxide of lithium, sodium, potassium, magnesium, calcium, barium, or strontium prior to contacting with the aluminum amalgam. Optionally. the quaternary ammonium bases are used for this neutralization, for example benzyltrimethylammonium hydroxide. The base is used in equivalent amount to the acid so that R; is replaced by the corresponding metal or quaternary ammonium cation. Alternatively, instead of the hydroxides or oxides, there are employed the hydrides, the carbonate, the bicarbonates, or the alkoxides, for example lithium hydride. potassium carbonate, sodium bicarbonate, magnesium methoxide, and the like. which form the corresponding Formula-Xla salts with the Formula-XI free acid. Alternatively, a metal or quaternary-ammonium carboxylate com pound or Formula Xla salt carried forward from the epoxidation step. whether isolated in that step or not, is

employed in the reductive step with aluminum amalgam. It is preferred that the Formula-XIa salt be soluble in the organic diluent-alkanol-water or organic diluentwater medium used for the reduction step. By using the above-described salts, the reduction step proceeds smoothly without formation of insoluble aluminum salts which hinder the reaction. Following the reduction or hydrolysis step, the R cations are replaced with hydrogen by means known in the art, for example by acidification and extraction of the acid compound into an organic phase.

The desired Formula-XII hydroxy products are iso' lated by filtration of the reaction mixture, advantageously after addition of magnesium sulfate as a filter aid, and evaporation of the organic diluents. The Formula-XII products are then hydrolyzed if desired to remove Si(A);; from C-15, and the [la and I15 products of Formula Xlll are separated, if desired, by procedures known in the art, e.g.. chromatography on silica gel.

The products of Formula-XII are all of the PGE -type and include PGE PGE l5-acetate, PGE methyl ester, PGE l5acetate methyl ester, POE and PGE methyl ester with an OSi--(A) at C- l 5, the corresponding 15B compounds, and compounds corresponding to all of those wherein hydroxy is attached to C11 in beta configuration.

As mentioned above, the transformation of X to XI to Xll usually gives a mixture of Formula-XII PGE'type products, part with alpha and part with beta configuration for the hydroxy at C-l 1. There are several alternatives regarding that mixture. If OSi(A) is attached to C-lS, that can readily be transformed by hydrolysis to -OH. These hydrolyses are carried out by prior art procedures known to be useful for transforming silyl ethers to alcohols. See, for example, Pierce, cited above, especially p. 447 thereof. A mixture of water and sufficient of a water-miscible organic diluent to give a homogeneous hydrolysis reaction mixture represents a suitable reaction medium. Additon of a catalytic amount of an organic or inorganic acid hastens the hydrolysis. The length of time required for the hydrolysis is determined in part by the hydrolysis temperature. With a mixture of water and methanol at 25 C., several hours is usually sufiicient for hydrolysis. At 0 C., several days is usually necessary. Also, if O- COCH is attached to C-lS, that can readily be transformed to ()H by acid-catalyzed alcoholysis as described above for removing the acetyl group of the Formula-VIII and -IX PGA-type starting materials. Both of those transformations are shown in Chart A, i.e., Xll to Xlll. Either before or after those transforma tions of Xll to Xlll, the Formula-XXI or -Xlll mixture of Ila and Ill? isomers can be separated by methods known in the art, advantageously by chromatography on silica gel.

Further regarding the Formula-XIII compounds, those compounds wherein the configuration of the hydroxy at C1] is beta are within the scope of the Formula-V novel compounds of this invention. and those compounds wherein the configuration of the hydroxy at C-] l is alpha and B is are within the scope of the Formula-VII novel compounds of this invention. Both groups of novel compounds are used for pharmacological purposes described above for those compounds. the acids also being useful as reactants to prepare pharmacologically useful esters and pharmacologically acceptable and useful salts, both as described above. Moreover, Formula-XIII compounds wherein the configuration of the hydroxy at Cl 1 is alpha and B is H OH are PGE and PGE methyl ester, both of known pharmacological utility.

Still further regarding the separated compounds of Formulas XII and XIII, when a compound with one configuration at C-1 1, either alpha or beta, is desired as an intermediate or for pharmacological purposes, the other isomer is readily dehydrated to give additional Formula-X PGA-type starting material which is then used as a starting material according to the processes defined in Chart A and procedures described above to give additional of the desired isomer. These dehydrations are accomplished by procedures known in the art for dehydration of PGE-type compounds to PGA-type compounds. See, for example, Pike et al., Proc. Nobel Symposium II, Stockholm (1966), lnterscience Publishers. New York, p. 162 I967), and British Specification l,()97,533. These are acidic dehydrations, and alkanoic acids of 2 to 6 carbon atoms, inclusive, especially acetic acid, are preferred for this purpose. Dilute aqueous solutions of mineral acid, e.g., hydrochloric acid, especially in the presence ofa solubilizing diluent,

e.g., tetrahydrofuran, are also useful as reagents for these acidic dehydrations, although these reagents may also cause partial hydrolysis of the Formula-XII or -Xlll methyl esters to carboxylic acids. A Si(A);i moiety at C is also removed during all of these acidic dehydrations.

Still further regarding the Formula-XII and -XIII compounds, either as mixtures or separately, any of those is transformed to other useful compounds or mixtures by changing these PGE-type compounds to PGF-type products by reducing the ring carbonyl at C9 to alpha hydroxy or beta hydroxy. Those transformations are shown in Chart B.

In Chart B, R is hydrogen, methyl, or -Si-(A) R is hydrogen or methyl, R is hydrogen or Si(A) and G is wherein R is hydrogen or -Si(A);; when R and R are hydrogen; R is hydrogen, acetyl, or Si(A);, when R; and R, are methyl; and R is Si(A):1When R, is Si(A) wherein A is as defined above, with the proviso that when R is Si(A) R is also Si- (A),-;. Further in Chart B, B in Formula XVI is and indicates attachment to the cyclopentane ring in alpha or beta position.

CHART B WQORZ Xll (fiilylation) (reduction) (hydrolysis or alcoholysis) Referring to Chart B, the starting material XII can be a mixture of compounds with regard to the configuration of Oil, or the starting material can be stereochemically pure with respect to C-1 1 depending upon whether there has been an earlier separation of Ila and 1 IB isomers (see above discussion of Chart A reactions).

The transformation of PGE-type starting material XII to PGF-type product XVI involves reduction of a ring carbonyl to a ring hydroxy. This process is known in the art for some of the compounds encompassed by Formula XII, i.e., when the configuration at C-l l is alpha and the configuration at C-15 is S. For the other com- 25 pounds encompassed by Formula XII, this reaction is novel, and novel Formula-XV and XVI compounds are produced.

For this carbonyl-to-hydroxy reduction, methods known in the art are used. See, for example, Pike et al., J. Org. Chem. 34, 3552 (I969). Use is made of any of the known ketonic carbonyl reducing agents which do not reduce ester or acid groups or carbon-carbon double bonds. Examples of those are the metal borohydrides, especially sodium, potassium, lithium, and zinc borohydrides, lithium (tri-tertbutoxy) aluminum hydride, metal trialkoxy borohydrides, e.g., sodium trimethoxyborohydride, and diisobutylaluminum hydride. The sodium, potassium, and zinc borohydrides are preferred for this reduction, especially zinc borohydride.

Unexpectedly, the amalgamated aluminum metal found useful above in transforming the Formula-XI epoxides to Formula-XII hydroxy compounds has also been found useful as an agent for this carbonyl-tohydroxy reduction of PGE-type compounds to PGF- type compounds. Either the PGE-type salts or the PCB- type esters are employed, for example the Formula-XII hydroxy compounds produced from the Formula-XI epoxides with or without intermediate isolation. Furthermore, the Formula-XI epoxides may be subjected to the combined epoxide-reduction and carbonyl reduction reactions practically simultaneously by operating at higher temperatures, for example 4060 C., although it is preferred for high yields of the 11- hydroxy compounds that the reductions be done stepwise. The solvents which are operable for this reduction are generally the same as those found useful for the epoxide-reduction step. Somewhat higher tempera tures or longer reaction times are required for the carbonyl-to-hydroxy reduction, however. For example, at 25 C., about 4 to 24 hours are required; at higher temperatures, e.g., 5()-60 C., about one to 2 hours are sufficient.

This carbonyl reduction usually produces a mixture of PGF type and PGF type compounds, i.e., compounds with the alpha configuration and compounds with the beta configuration for the hydroxy at C-9. This mixture of alpha and beta isomers is separated by methods known in the art, e.g., chromatography on silica gel. See Pike et al., ibid., for example. If the Formula-XII starting material is a mixture of l l a and 11B isomers, then this reduction will usually produce four isomers, i.e., 901,1 1a, 901,1 1B,9/3,1 1a, and 93,113. Those compounds are also separated from such mixtures by silica gel chromatography.

Regarding the transformation of XII to XIV in Chart B, it will be observed that the parameters for XII are such that all XII compounds are included in XIV. In other words. the transformation XII to XIV is an optional process step in proceeding from XII to XV. The reason for this is as follows. During the reduction of XIV to XV, the ratio of 9a-hydroxy and QB-hydroxy compounds formed will be different when R in XIV is hydrogen than when R-, is Si(A) For example, with the Formula-XIV compound wherein R, is hydrogen. G is H OH.

26 and R O- represents HO, i.e.. Ila-hydroxy, sodium borohydride reduction gives 42 parts of the corresponding Formula-XV Jo-hydroxy compound. and 58 parts of the 9B-hydroxy compound. But with the Formula-XIV compound wherein R is hydrogen, G is H o -siacHni,

and R O- represents (CH );,SiO-, sodium borohydride reduction gives parts of the corresponding Formula-XV 9a-hydroxy compound and 15 parts of the 9B-hydroxy compound. Similar differences are observed with the other isomers encompassed by Formula XIV although not necessarily in the same direction. Accordingly, whether R in Formula XIV is to be hydrogen or Si(A);; depends on the particular formula XV C-9 isomer desired and the influence of silylation on the isomer ratio. For any particular Formula-XIV starting material, the latter is readily determined by small scale reduction with and without silylation. When silylation before carbonyl reduction is indicated, largely for economic reasons, it is preferred that A be methyl, i.e., that R be (CH -,);,Si-.

This transformation of XII to XIV wherein R is Si- (A)=. is carried out as described above for the transformation of hydroxy to OSi-(A) at C-l5 prior to the Chart A reactions. When R in XII is hydrogen, the COOH is also transformed in part or entirely to COOSi(A) with prolonged silylation and excess silylating agent. It is optional in transforming XII to XIV wherein R is Si(A),-, whether or not the COOH of XII is esterified to COOSI(A):. When G in Formula XIV is H' OHorH 'oH,

those OH are also transformed to OSI(AI3 by this silylation.

With regard to the Formula-XV carbonyl reduction product (Chan B), when the method used to isolate said product does not remove any Si-(Ah groups which may be present, that is accomplished as described above for the removal of Si-(AL; groups from Formula-XII products (Chart A, XII to XIII). Also, when G in Formula XV is H OCOCH-I or H ococm,

the acetyl is removed by alcoholysis also as described above for changing acetoxy at C-15 to hydroxy. These reactions are shown in Chart B as XV ro XVI.

When R in Formula XVI is methyl and the compound wherein R is hydrogen is desired, that methyl ester is saponified, by methods known in the art. See, for example, Just et al., .I. Am. Chem. Soc. 91, 5371 (1969). This saponification also changes a C-l5 acetate to a C-15 hydroxy.

The compounds encompassed by Formula XVI include the known compounds PGF- PGF and the methyl esters of those. Also included in ormula XVI are the novel compounds ISB-PGF 15B- PGF and the methyl esters of those. All of these new and old compounds are l [oz-hydroxy compounds. Also included in Formula XVI are the corresponding 27 but novel I lB-hydroxy compounds which are also encompassed by Formula V and which are useful for the pharmacological purposes described above either as such or transformed into salts or esters as described above.

When one of these Formula-XVI compounds has the R or epi configuration for the hydroxy at C- l 5, and the corresponding compound with the S configuration at (-15 is desired, or when one of these Formula-XVI compounds has the S configuration for the hydroxy at G15, and the corresponding compound with the R or epi configuration at C- l is desired, those desired compounds are made by the processes of Chart C. In Chart C, R R R B, and are as defined above.

The overall process scheme of Chart C is to start with one particular C-l5 isomer of a compound encompassed by Formula XVI, i.e., either (S) or l5(R). The C-15 hydroxy of that isomer is oxidized to a ketonic carbonyl (XVII). Then, after an optional silylation of the C 9 and C-1 1 hydroxy groups (XVIII), the C-l5 carbonyl is reduced back to a secondary hydroxy group. That reduction produces two C-l5 hydroxy isomers, one with S configuration and one with R or epi configuration. After removal of any silyl groups, the isomers XIX and XX are separated. One of the isomers will be the same compound used as starting material (XVI) The other isomer will be the desired product. The starting material isomer is recycled to produce more of the desired isomer This reaction scheme has previously been used to transform PGF to I58- PGF l See Pike et al., J. Org. Chem. 34, 3552 (I96 Referring now to Chart C, starting material XVI (from Chart B) is a single compound, a mixture of two compounds, one with alpha and one with beta configuration at C-9, or a mixture of four compounds, i.e., 9a,l la,9a,l 1393,: la, and 95,115.

CHARTC 'WA XV' (Oxidation) DORE XVII (si lylationl R. XVIII R7 Q/\/\/ J (reduction) i lhydrolysi s) XIX For the oxidation of XVI to XVII, any oxidizing agent can be used which will oxidize an allylic alcohol to an a,B-unsaturated ketone or aldehyde. Examples of those are 2,3-dichloro-5,6-dicyano-l,4-benzoquinone, activated manganese dioxide, or nickel peroxide (see Fieser et al., Reagents for Organic Syntheses, John Wiley & Sons, Inc, New York, N.Y., I967, pp. 215, 637, and 73 I Alternatively, these oxidations are carried out by oxygenation in the presence of the 15- hydroxyprostaglandin dehydrogenase of swine lung (see Arkiv for Kemi 25, 293 (I966)). These reagents are used according to procedures known in the art. See, for example, J. Biol. Chem. 239, 4097 I964).

Regarding the transformation of XVII to XVIII in Chart C, it will be observed that the parameters for XVIII are such that all XVII compounds are included in XVIII. In other words, the transformation of XVII to XVIII is an optional process step in proceeding from XVII to XIX and XX. The reason for this is as follows. During the reduction of XVIII to XIX and XX, the ratio of XIX to XX obtained will be different when R in XVIII is hydrogen than when R is Si(A) For example, reduction of the Formula-XVIII 901,1 la-isomer wherein R and R are both hydrogen with zinc borohydride gives the corresponding Formula-XIX and -XX in the amounts 43 parts of XIX (R or epi configuration) and 57 parts of XX (S configuration). On the other hand, when R in the Formula-XVIII reactant is -Si(A the amounts with the same reducing agent are 27 parts of XIX and 73 parts of XX. Similar differences are observed with the other isomers encompassed by Formula XVII; although not necessarily in the same direction. Accordingly, whether R in formula XVIII is to be hydrogen or Si(A) epends on the particular C-l5 isomer desired and the influence of silylation on the isomer ratio. For any particular Formula-XVII starting material, the latter is readily deter mined by small scale reductions with and without silylation. When silylation before carbonyl reduction is indicated, largely for economic reasons, it is preferred that A be methyl, i.e., that R be (CH Si.

These silylations are carried out described above for the Chart A and Chart B silylation.

The carbonyl reduction of XVIII to XIX is carried out as described above for the transformation of PGE type Formula-XIV compounds to PGF-type Formula- XV compounds. As for those reductions, the sodium, potassium and zinc borohydrides are preferred as reducing agents, especially zinc borohydride.

When the method used to isolate the carbonyl reduction product does not remove any Si (A);, groups which may be present, that is accomplished as de 29 scribed above for the removal of -Si(A);, groups from Formula-XII products (Chart A, XII to XIII).

The Formula-XIX and -XX products are separated from each other by methods known in the art, for example, silica gel chromatography. See, for example, Pike et al., J. Org. Chem, 34, 3552 I969) for this type of separation.

If one of the isomers or isomer mixtures of Formulas XIX or XX is not desired for a pharmacological use as such or transformed to esters or pharmacologically acceptable salts described above, that isomer or isomer mixture is recycled as a Formula-XVI starting material in the processes of Chart C to produce additional of the desired isomer.

The products of Formulas XIX and XX wherein the configuration of the C11 hydroxy is beta are encompassed by Formula V. The products of Formula XIX wherein the configuration of the C-1 1 hydroxy is alpha are encompassed by Formula VI. The intermediates of Formula XVII are encompassed by Formula VII. Thus, all of the compounds are useful for the pharmacological purposes described above for the Formula V, VI and VII compounds. The compounds prepared as in Chart C are also useful to make the other esters and the pharmacologically acceptable salts of the Formula V, VI and VII compounds also as described above.

There are two particular embodiments of the novel process of this invention which are especially preferred. One of those embodiments provides an optional route to PGF and starts with lB-PGA acetate methyl ester, the most abundant component of Plexuura lwmomalla (Esper), I792, forma R. The other embodiment provides a preferred route of PGE and starts with PGA readily obtained as described above by maintaining colonies or colony pieces of Plexaura homomallu (Esper), I792, forma S in contact with water in a temperature range up to 50 C. until substantially free of PGA acetate methyl ester.

The first of these embodiments is shown in Chart D, and the second is shown in Chart E. All of these Chart D and Chart E reactions and reagents for effecting them are described generically and specifically above, and all are exemplified below. In Charts D and E, it is preferred that Si(A)- be SiCH;,),-,. Also in Charts D and E, it is optional whether silylation of l5-oxo-PGF (Chart D) or PGA (Chart E) produces the corresponding Si(A) ester-ether or only the ether.

CHART D ISBPGA, acetate methyl ester (oxidation) l5fi-P(EA. acetate methyl ester (1 and B l(),l lepoxidcs (reduction) lS/i-PGF; and l ltil SB-PGE l5-acetate methyl esters (separation) ISB-PGE Y l5-acetate methyl ester lullylatam) ISBPGE ll-Si-(A), ether l5 acetate methyl ester (reduction, hydrolysis) ISIS-POP and l5B-PGF. IS-acctatc methyl esters Mscparation) CHART E PGAQ l, (silylation) PGA, lS-Si-(Al ether 1 (oxidation) PGA IS-Si-(Ah ether [3 and a lU.l l-epoxides l (reduction, hydrolysis) PGE and l I01 PGF 1 (separation) Po E,

The invention is more fully understood by the following Examples and Preparations All temperatures are in degrees centigrade.

Ultraviolet spectra are recorded on a Cary Model 15 spectrophotometer.

The collection of chromatographic eluate fractions starts when the eluant front reaches the bottom of the column.

Brine, herein, refers to an aqueous saturated sodium chloride solution.

The AIX solvent system used in thin layer chromatography (TLC) is made up from ethyl acetate-acetic acid-2,2,4-trimethylpentane-water (:20:50:l00) according to M. Hamberg and B. Samuelsson, J. Biol. Chem. 24], 257 (I966).

PREPARATION I To distinguish Plexaura lwmomalla (Esper), I792, forma R from Plexuura humomalla (Esper), I792, forma S, a TLC method is used. A specimen approximately 2 cm. in length is harvested and placed in a small vial, with a small amount of water if necessary to insure it is wet, and kept closed for 624 hrs. About one ml. of methanol is then added and the sample is either shaken for 2 hrs. at about 25 C. or is stored for l6-24 hrs. at about 10 C. A sample of the liquid l()2l A) is spotted on a TLC plate. It is preferred to use a fluorescent-treated silica gel plate, e.g. Uniplate Silica Gel GF (Analtech, Inc., Newark, Del). As reference standards, spots of PGA and l5/3-PGA are also applied. The plate is developed in the A-IX system (Hamberg and Samuelsson, J. Biol. Chem. 24l, 257 1965)). The spots are finally visualized with vanillin-phosphoric acid spray (McAleer, Arch. Biochem. E. Biophys. 66. I20 1957)). Comparison of the unknown with the two reference spots is then made and the identity of the coral established (forma S corresponding to PGA- forma R to ISB-PGA PREPARATION 2 PGA-g from Plexauru lwmumullu (Esper), I792 forma S Colonies of Plexauru lwmomalla (Esper). 1792, forma S, collected from reefs off the north shore of Jamaica, are frozen by contact with solid carbon dioxide within one hour after removal from the reef waters. The frozen colonies are maintained in insulated boxes containing solid carbon dioxide (temperature below about 20 C.) until ready for thawing. Then, the frozen colonies (700 g.) are ground to a small particle size (Waring blender) and mixed with 1500 ml. of water. The mixture is maintained about 20 hrs. at about 25C. with stirring. Then, the mixture is filtered through a pad ofdiatomaceous earth. and the filtrate is acidified with concentrated hydrochloric acid to pH about 2-3. The acidified filtrate is extracted four times with ethyl acetate. The extracts are combined, filtered, washed with brine, dried with anhydrous sodium sulfate, and evaporated under reduced pressure to give 1 l g. of oily residue.

The solid residue on the diatomaceous earth filter pad is stirred 2 hours in methanol (enough to cover said residue) at 25 C. The mixture is then filtered, and the filtrate is evaporated to give 14 g. of oily residue.

The two oily residues are combined and chromatographed on 1500 g. of aeid washed silica gel, eluting successively with 8 l. of a 25 to 65% gradient of ethyl acetate in Skellysolve B, 8 l. of a 65 to l gradient of ethyl acetate in Skellysolve B, and 5 l. of 2% methanol in ethyl acetate. collecting 500 ml. fractions. (Skellysolve B is a mixture of isomeric hexanes). Fractions 8l2 are combined and evaporated to give a small amount of PGA containing a trace of PGA methyl ester. Fractions -18 are combined and evaporated to give 9.54 g. of PGA.,;. Fractions 35-40 are combined to give 0.4 l4 g. of PGE PREPARATION 3 ISB-PGA from Plexaura Homomalla (Esper), 1792, forma R Colonies of Ilatuuru homumullu (Esper), l792, forma R, collected from reefs off the southeast shore of Florida near Miami, are chopped into chunks several inches long. The chunks are frozen by contact with solid carbon dioxide with one hour after removal from the reef waters. The frozen colony pieces are maintained in insulated boxes containing solid carbon dioxide (temperature below about C.) until ready for thawing. Then, colony pieces ((100 g.) are mixed with I500 ml. of water. The mixture is stirred and maintained at C. for 23 hours. The mixture is then filtered through a pad of diatomaceous earth, and the filtrate is acidified to pH about 2-3 with concentrated hydrochloric acid. The acidified filtrate is extracted four times with ethyl acetate. The extracts are combined. filtered, washed with brine. dried with why drous sodium sultate, and evaporated to give 9.2 g. of oily residue.

The solid residue on the diatomaceous earth pad is stirred [5 hours in methanol (enough to cover said residue) at 25 C. The mixture is then filtered, and the filtrate is evaporated. The residue is dissolved in ethyl acetate, and the solution washed successively with 3 N hydrochloric acid and brine, dried with anhydrous sodium sulfate, and evaporated to give 5.83 g. of an oily residue.

The second oily residue and 8.2 g. of the first oily residue are combined and chromatographed on one kg. of acid-washed silica gel, eluting successively with 3-l. portions of 25%, 35%, 45%,559/0, and 65% ethyl acetate in Skellysolve B, collecting 500-ml. fractions, Fractions 18-22 are combined and evaporated to give 5.54 g. of l5B-PGA Fractions l5l7 are combined and evaporated to give 1.37 g. of lSB-PGA methyl ester.

PREPARATION 4 PGA compounds from P/aruuru humumul/a (Esper), 1792, forma S Frozen colonies of Plexaura homumallu (Esper), I792, forma S (see Preparation 2) are broken manually into pieces several cm. in length. The pieces (500 g) are then covered with methanol and the mixture is maintained for 3 hours at 25 C. The mixture is then ground in a Waring blender and filtered, and the filtrate is evaporated under reduced pressure. The residue is dissolved in ethyl acetate, and the solution is washed successively with one N hydrochloric acid, water, and brine, dried with anhydrous sodium sulfate, and evaporated under reduced pressure. The oily residue is chromatographed on 2 kg. of acid-washed silica gel wet packed with Skellysolve B (a mixture of isomeric hexanes), eluting with 24 l. of a 25 to l0071 ethyl acetate in Skellysolve B gradient. The fractions which contain PGA acetate methyl ester, PGA acetate, PGA: methyl ester, and PGA shown by TLC with the A-IX system are separately combined and evaporated to give those compounds.

PREPARATION 5 l5B-PGA compounds from Plexauru humomaI/a (Esper), 1792, forma R Colonies of Plat-aura honwmalla (Esper), l792, forma R, collected from reefs off the southeast shore of Florida near Miami. are chopped into chunks several inches long. The chunks are frozen by contact with solid carbon dioxide within one hour after removal from the reef waters. The frozen colony pieces are maintained in insulated boxes containing solid carbon dioxide (temperature below about 20 C.) until the time for extraction. Then, the frozen colony pieces are ground to a small particle sice (Mitts and Merrill hogger; average largest dimension about 5 mm). The particles (I500 g.) are then stirred at high speed with 5 gallons of dichloromethane for 20 minutes at about 25 C. external temperature. The mixture of dichloromethane and particles is then filtered through a pad of diatomaceous earth, and the filtrate is evaporated to about a 2-liter volume at 30 C. under reduced pressure. The liquid which remains is washed with water, dried with sodium sulfate, and evaporated at 30 C. under reduced pressure.

The oily residue (60 g.) is chromatographed on 3 kg. of silica gel wet packed in Skellysolve B (a mixture of isomeric hexanes), eluting successively with a gradient of4 l. of Skellysolve B and 4 l. of 20% ethyl acetate in Skellysolve B, 27 l. of 20%, 18 l. of 50%, and 8 l. of ethyl acetate in Skellysolve B. collecting 600-ml. fractions. Fractions 394%) are combined and evaporated to give 24.3 g. of l5-PGA- acetate methyl ester. Between fractions (10 and 74 those fractions shown by TLC to contain l5f3--P(1A acetate are combined and evaporated to yield that compound. Fractions 74-7fi are combined and evaporated to give L03 g. of 153- PGA methyl ester. Fractions 83-9l are combined and evaporated to give 1.08 g. of lSB-PGE lS-acetate methyl ester. Still later fractions shown by TLC to contain [SB-POE methyl ester are combined and evaporated to yield that compound.

Detection of the respective compound by TLC is done by methods known in the art, e.g. by spotting the extract fractions on a TLC silica gel plate alongside 33 spots of the authentic compounds, developing the plate with the A -lX system, and observing which spots of the extract fractions correspond exactly to the spots of the authentic compounds.

Following the procedures of Preparation 5, but substituting Plexaura liomnmalla (Esper), 1792, forma S for the Plexaura homomulla (Esper), 1792, forma R of that example, there are obtained the corresponding compounds of 15(S) configuration, viz.: PGA acetate methyl ester, PGA acetate, PGA methyl ester, and PGE l5-acetate methyl ester, and PGE methyl ester.

PREPARATION 6 PGA and 5,6-trans-PGA Separation of PGA from 5,6-trans-PGA is done on a chromatographic column using a silver-saturated ion-exchange resin. Preferably a macroreticular ion exchange resin is used, e.g. a sulfonated styrene-divinylbenzene copolymer having surface area of 40-50 sq. m./g., 30-40% porosity. and total exchange capacity of 4.5-5.0 meg. per gram of dry resin, for example Amberlyst 15, available from Rohm and Haas Co., Philadelphia, Pa. The acid-form resin is packed in a column, washed with warm water, and converted to the silver fonn by passing a 10% silver nitrate solution through the column until the effluent shows a pH of 3.5-4.0. The column is then washed with water to remove ionic silver, and finally with denatured ethanol (Type 3A). A solution of a mixture of PGA and 5,6-trans-PGA e.g. fractions -18 of preparation 2, in ethanol is charged to the column. Elution with 3A alcohol then yields fractions which are combined according to their content of 5,6-trans-PGA (faster-eluting) or PGA Testing for the presence of 5,6-trans-PGA or PGA in the eluate is conveniently done by TLC using silver nitratetreated silica gel plates (e. g. Analtech Uniplates dipped in saturated ethanolic silver nitrate and dried) and developing with the AJX system. R, of PGA is 0.45; R of 5,6-trans-PGA 2 is 0.50. Combined fractions are concentrated partitioned between dichloromethane and a little water, dried over sodium sulfate, and concentrated under reduced pressure to yield the title compounds.

For quantitatively assaying the 5,6-trans-PGA content of mixtures of PGA and 5,6-trans-PGA a combination thinlayer-spectrophotometric assay is used. Silica gel-impregnated glass microfiber sheets (e.g. ITLC sheets of the Gelman Instrument Co., Ann Arbor, Mich.) are impregnaged with silver nitrate, using 5% ethanolic silver nitrate and drying. Spots of 100 to 200 pg of the PGA: mixture are applied and developed in the solvent system 2,2,4-trimethylpentane:ethy1 acetate: acetic acid: water (100:35:8:10, upper phase). The sheet is dried and sprayed with Rhodamine 6G (Applied Science Co., State College, Pa) and viewed under ultraviolet light. The areas containing the cis and trans materials (R of PGA 0.6: R;,,,5,6-trans-PGA 0.7) are marked, then exicised and eluted with methanol 1.9 ml.) and potassium hydroxide solution (0.1 ml. of 45%). After incubation at 40 for 30 min., the respective solutions are centrifuged and analyzed spectrophotometricaily at 278 nm.

Following the procedure of Preparation 6,5,6-trans- 15,8-PGA is separated from l5B-PGA PREPARATION 7 PGE and 5,6-trans-PGE Following the procedure of Preparation 6, PGE is separated from 5,6-transPGE as follows. A solution of a mixture of PGE and 5,6-trans-PGE is charged to the column. Elution with 3A alcohol yields fractions which are combined according to their content of 5,6-trans- PGE (faster eluting) or PGE Assay for 5,6-trans- PGE or PGE is done by TLC as for the PGA- -type compounds above. R, of PGE is 0.13; R, of 5,6-trans- PGE is 0.17. Combined fractions are concentrated, dried over sodium sulfate, and concentrated under reduced pressure to yield the title compounds.

PREPARATlON 8 PGA lS-Acetate Methyl Ester, separation from 5,6-Trans-PGA l5-Acetate Methyl Ester A mixture of PGA 15-acetate methyl ester and 5,6- trans-PGA l5-acetate methyl ester (1 1.0 g., :15) is dissolved in 415 ml. of a solution of methanol-wateracetic acid (-5-04) and mercuric acetate (6.1 g.), and left standing at about 25 C. for 30 min. Water (250ml.) is added and the mixture extracted twice with 700 ml. of Skellysolve B. The Skellysolve B phase is washed with ml. of 60% methanol, dried over sodium sulfate, and concentrated to an oil (4.35 g.) having a high content of 5,6-trans-PGA l5-acetate methyl ester. The aqueous methanol phase is acidified with 32 ml. of 6 N. hydrochloric acid and the mixture is extracted with two portions of 700 ml. of Skellysolve B. The organic phase is dried over sodium sulfate and concentrated to an oil (5.53 g.) This last material is subjected to the same procedures again, using 350 m1. of the methano1water-acetic acid and 4.6 g. of mercuric acetate. There is recovered from the work-up of the aqueous-methanol phase a fraction (3.92 g.) of the title compound containing only a small percentage of the 5,6-trans-PGA compound.

Following the procedure of Preparation 8,5,6-transl5B-PGA l5-acetate methyl ester is separated from l5B-PGA l5-acetate methyl ester.

1n the following examples, the above-described 5,6- tran -P and 5,6trans-l5B-PG compounds are subjected to the same transformations as the PG and 15B- PG compounds disclosed herein and illustrated hereafter.

EXAMPLE 1 l5B-PGA Methyl Ester A solution of 70% aqueous perchloric acid (50 drops) in 50 drops of water is added to a solution of l5B-PGA acetate methyl ester (2.0 g.) in 100 ml. of methanol. The mixture is stirred for 15 hours at 25 C. and then diluted with 80 ml. of water. The methanol is removed under reduced pressure, and the aqueous residue is extracted with ethyl acetate. The extract is washed successively with water and brine, dried with anhydrous sodium sulfate, and evaporated. The residue is chromatographed on 200 g. of silica gel, eluting with 2.5 l. ofgradient of 2070% ethyl acetate in Skellysolve B (a mixture of isomeric hexanes), collecting l00-ml. fractions. Fractions 15-19 are combined and evaporated to give 727 mg. of lSB-PGA- methyl ester.

EXAMPLE 2 PGA Methyl Ester A solution of 15B-PGA methyl ester (250 mg.) in 20 ml. of anhydrous tetrahydrofuran is cooled to C. in an atmosphere of nitrogen. Tributylamine (1.5 ml.) is added, and the mixture is stirred at 0 C. while adding methanesulfonyl chloride (1 ml.) dropwise. The mixture is stirred 30 minutes at 0 C. Then, ml. of water is added, and the mixture is allowed to warm to 25 C. and is stirred for one hour. The tetrahydrofuran is evaporated under reduced pressure, and the aqueous residue is extracted with ethyl acetate. The extract is washed successively with one N hydrochloric acid, water, and brine, dried with anhydrous sodium sulfate, and evaporated. The residue is chromatographed on 30 g. of silica gel, eluting with 800 ml. of a gradient of 70% ethyl acetate in Skellysolve B, collecting ml. fractions. Fractions 14-16 are combined and evaph orated to give 58 mg. of PGA methyl ester. Fractions 12 and 13 are combined to give 49 mg. of the starting material, l5/3PGA methyl ester Following the procedure of Example 2, PGA methyl ester is transformed to a mixture of PGA and 15B- PGA- methyl esters, the two compounds being separated as in Example 2.

EXAMPLE 3 PGA l5-formate and lSB-PGA 15-formate A solution of sodium carbonate (50 mg.) in 7.5 ml. of anhydrous formic acid is added to PGA (0.25 g.) This mixture is stirred under nitrogen at 25C. for 2 hrs. The reaction mixture is concentrated under reduced pressure, taken up in benzene, and again concentrated under reduced pressure. The residue is chromatographed on acid-washed silica gel (e.g. Mallinckrodt Silicar CC-4), eluting with a gradient of 2575% ethyl acetate-skellysolve B (isomeric hexane mixture) and collecting fractions. Those fractions shown by TLC to contain the respective l5-formate compound, separated from its isomer and free of starting material and impurities, are combined and concentrated under re duced pressure to give the title compounds.

EXAMPLE 4 PGA and 15B-PGA PGA l5-formate (100 mg., Example 3) is dissolved in a mixture of 10 ml. of methanol and 25 ml. of saturated aqueous sodium bicarbonate solution. The solution is stirred under nitrogen at 25 C. for 2.5 hrs. then 5 ml. of water and 2 ml. of l N. hydrochloric acid are added, and the solution concentrated. The aqueous residue is adjusted to pH 2-3 and extracted three times with ethyl acetate. The combined extracts are washed with water. dried over sodium sulfate, and concentrated to yield PGA Similarly,, hydrolysis of ISB-PGA 15-formate (Example 3) yields l5B-PGA EXAMPLE 5 15B-PGA 10,1 l-Epoxide Acetate Methyl Ester Hydrogen peroxide (350 ml.; aqueous) is added with stirring to a solution of 15B-PGA acetate methyl ester (265 g.) in 5000 ml. of methanol under a nitroge atmosphere at 20 C. Then, one N aqueous potassium hydroxide solution (50 ml.) is added gradually during one hour with stirring at 20C. The mixture is stirred an additional 2 hours at 20C. Then, one N hydrochloric acid ml.) is added, and the methanol is removed under reduced pressure at 35C. The residue is dissolved in 3000 ml. of ethyl acetate, and the solution is washed 3 times with 500-ml. portions of water. The combined water washes are extracted with 300 ml. of ethyl acetate. The two ethyl acetate solutions are combined, washed with brine, dried with anhydrous sodium sulfate and evaporated to give 2.75 g. ofa mixture of the alpha and beta 10,] l-epoxides of lSB-PGA acetate methyl ester.

EXAMPLE 6 PGA l0,l l-Epoxide Methyl Ester Hydrogen peroxide (0.3 ml.; 30% aqueous) and one N aqueous sodium hydroxide (0.5 ml.) are added to a solution of PGA methyl ester (229 mg.) in 10 ml. of isopropyl alcohol at 0C. After 2.5 hours at 0C., 10 ml. of water and one ml. of one N hydrochloric acid are added, and the isopropyl alcohol is removed under reduced pressure. The residue is extracted with ethyl acetate. The extract is washed successively with water and brine, dried with anhydrous sodium sulfate, and evaporated. The residue is chromatographed on 30 g. of silica gel, eluting with 800 ml. of a gradient of 20-70% ethyl acetate in Skellysolve B, collecting 25- ml. fractions. Fractions 5-10 are combined and evaporated to give 136 mg. of a mixture of the alpha and beta l0,l l-epoxides of PGA methyl ester.

EXAMPLE 7 PGA Acetate Methyl Ester 01 and B 10,] l-Epoxides Refer to Chart A.

A solution of PGA l5acetate methyl ester (1.954 g.) in 30 ml. of dimethoxiyethane (DME) is cooled to 55C. under nitrogen, and 5.25 ml. of t-butyl hydroperoxide is added. Then, 5 ml. of 0.25 N. methanolic lithium hydroxide (prepared from the mono-hydrate) is added over min. After about 46 hrs. an additonal 2.5 ml. of the base is added over 50 min. Finally, after about 23.5 hrs. the reaction is complete, shown by TLC. The mixture is acidified to pH 56 with l N. hydrochloric acid and is concentrated under reduced pressure. The residue is taken up in ethyl acetate, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The product, 2.0 g., contains the title alpha and beta compounds in a ratio of 6:], respectively, as shown by gas chromatography.

Following the procedures of Example 7, but replac ing the lithium hydroxide solution with metranolic magnesium methoxide (prepared from magnesium and anhydrous methanol), there is obtained a product containing the alpha and beta epoxides in a ratio of 4:1.

Following the procedures of Example 7, but replacing the DME with a mixture of toluene-DME (10:1) and replacing the lithium hydroxide with Triton B (benzyltrimethylammonium hydroxide) in methanol, there are obtained the alpha and beta epoxides in a ratio of 7.211.

Following the procedures of Example 7, but replacing the DME with a mixture of tolueneDME 1:1 and holding the reaction temperature at 40 C, the prod uct contains the alpha and beta epoxides in a ratio of 6.2:1.

Following the procedures of Example 7. but replacing the DME with tetrahydrofuran (THF) and replac- 37 ing the lithium hydroxide solution with Triton B, there are obtained the alpha and beta epoxides in a ratio of 4.5: 1.

EXAMPLE 8 lB-PGE 15-Acetate Methyl Ester and 1 1B, ISB-PGE lS-Acetate Methyl Ester Granular aluminum metal (50 G.) is added to a solu tion of mercuric chloride (50 g.) in 2 l. of water. The mixture is swirled until hydrogen gas evolution starts to become vigorous (about 30 seconds). Then, most of the aqueous solution is decanted, and the rest is removed by rapid filtration. the amalgamated aluminum is washed rapidly and successively with two 200ml. portions of methanol and two ZOO-ml. portions of anhydrous diethyl ether. The amalgamated aluminum is then covered with anhydrous diethyl ether until used.

Methanol (250 ml.) and water (25 ml.) are added to a solution of a mixture of the alpha and beta 10,1 lepoxides of lSB-PGA acetate methyl ester (275 g.) in 2500 ml. of diethyl ether. the mixture is cooled to C. and the amalgamated aluminum prepared as above from 50 g. of aluminum metal is added. The mixture is stirred and maintained at about 25C. with external cooling. After one hour, amalgamated aluminum prepared as above from 50 g. of aluminum metal is added. After an additional hour of stirring at 25 C., amalgamated aluminum prepared as above from 50 g. of aluminum metal and also 25 ml. of water are added. After an additional hour of stirring at 25 C., 200 g. of magnesium sulfate is added as a filter aid, and the mixture is filtered. The filter cake is washed throughly with dichloromethane, and the combined filtrate and washing are evaporated at 25 C. under reduced pressure to give a mixture 247 g.) of l5B-PGE IS-acetate methyl ester and l 1 B,15B-PGE l5-acetate methyl ester.

Part of this mixture (210 g.) is chromatographed on 30 kg. of silica gel wet-packed with 60 l. of 25% ethyl acetate in Skellysolve B (6-inch diameter column), eluting successively with 60-1. portions of 25%, 30%, 35% 40%, 45, 50%, 55%, and 60% ethyl acetate in Skellysolve B, collecting 4-l. fractions. Fractions 71-76 are combined and evaporated to give 27 g. of l 15,15,6- PGE l5-acetate methyl ester. Fractions 81-98 are combined and evaporated to give 115 g. of ISB-PGE IS-acetate methyl ester.

EXAMPLE 9 ISB-PGE l5-Acetate Methyl Ester and l l[5',15fi]-PC|E -Acetate Methyl Ester Anhydrous sodium acetate (0.5 g.) and zinc dust (500 mg.) are added to a solution of a mixture of the alpha and beta epoxides of l5/3-PGA acetate methyl ester. prepared in Example 5, in 5 ml. of acetic acid. This mixture is stirred at 25C. in an atmosphere of nitrogen and cooled to about 15 C. One-half ml. of a solution of chromium (Ill) chloride hexahydrate (300 mg.) in 1 ml. of water is added, and the mixture is stirred at 0C. for 3 hours. The mixture is then diluted with ethyl acetate. and the solution is washed successively with four portions of water, one N hydrochloric acid. sodium bicarbonate solution. and brine, dried with anhydrous sodium sulfate, and evaporated. The residue is chromatographed on g. of silica gel, elut ing with (100 ml. ofa gradient of 20-75% ethyl acetate in Skellysolve B, collecting 2.5-ml. fractions. Fractions l0 and l l are combined to give l 18,158 PGE lS-acetate methyl ester. Fractions 13 and 14 are combined to give 15B-PGE l5-acetate methyl ester.

EXAMPLE 10 FOB; Methyl Ester and l l,B-PGE Methyl Ester Freshly prepared chromium (ll) acetate (450 mg., argon atmosphere; Inorganic Syntheses, 8, is added to a solution of 136 mg. of epoxides (Example 6) in a mixture of 3 ml. of acetic acid and one ml. of water in an atmosphere of argon at 0 C. The mixture is stirred at 5 C. under argon for 18 hours. Ice is then added to the mixture, and that mixture is extracted with ethyl acetate. The extract is washed successively with water, one N hydrochloric acid, sodium bicarbonate solution, and brine, dried with anhydrous sodium sulfate, and evaporated. The residue is chromatographed on silica gel (20 g.), eluting with 600 ml. of a gradient of 20-100% ethyl acetate in Skellysolve B. collecting 20 ml. fractions. Fractions 19-22 are combined and evaporated to give 27 mg. of l lB-PGE: methyl ester. Fractions 24-27 are combined and evaporated to give 5 mg. of PCB; methyl ester.

EXAMPLE 1 l PGE and 1 1B-PGE Hydrogen peroxide (0.35 ml. 30% aqueous) is added to a solution of PGA; (200 mg.) in 5 ml. of methanol. The mixture is cooled to 20 C., and 0.75 ml. of one N aqueous sodium hydroxide solution is slowly added with stirring. After one hour of stirring at 20C., one ml. of one N hydrochloric acid is added. and the mixture is evaporated under reduced pressure. The residue is dissolved in ethyl acetate, and the resulting solution is washed successively with water and brine, dried with anhydrous sodium sulfate, and evapo rated. The residue is treated with amalgamated aluminum as described in Example 8, using 2.5 ml. diethyl ether, 0.25 ml. methanol, and 0.03 ml. water. the amalgamated aluminum being added in 2 portions. When the reduction is complete, ethyl acetate and one N hydrochloric acid are added to the reaction mixture and the mixture is separated in a separatory funnel. The ethyl acetate layer is washed successively with one N hydrochloric acid, water, and brine, dried with anhy' drous sodium sulfate, and evaporated. The residue is subjected to preparative thin layer chromatography to give PGE and l 1B-PGE in the ratio 1:2.

EXAMPLE [2 15BPGE2 and 1113,15/3-PGE2 Following the procedure of Example I l. 15B-PGA is transformed to lSB-PGEz and llB,l5,8-PGE those being obtained in the ratio 1:1.

EXAMPLE l3 PGE- and 1 lB-PGE Hexamethyldisilizane l ml.) and trimethylchlorosilane (0.2 ml.) are added with stirring to a solution of PGA (250 mg.) in 4 ml. of tetrahydrofuran at 0 C. under nitrogen. This mixture is maintained at 5 C. for 15 hours. The mixture is then evaporated under re duced pressure. Toluene is added and evaporated twice. Then the residue is dissolved in 6 ml. of methanol. and the solution is cooled to 20 C. Hydrogen peroxide (0.45 ml.; 30% aqueous) is added. Then, one N sodium hydroxide solution (0.9 ml.) is added drop- 

1. A COMPOUND OF THE FORMULA:
 2. A compound of the formula:
 3. A compound according to claim 2 wherein the hydroxy is attached to the ring in alpha configuration.
 4. A compound of the formula: 